In vitro Reconstitution and Imaging of Microtubule Dynamics by Fluorescence and Label-free Microscopy

William Graham Hirst, Christine Kiefer, Mohammad Kazem Abdosamadi, Erik Schäffer, Simone Reber

STAR Protoc. 2020, Nov 24


Dynamic microtubules are essential for many processes in the lives of eukaryotic cells. To study and understand the mechanisms of microtubule dynamics and regulation, in vitro reconstitution with purified components has proven a vital approach. Imaging microtubule dynamics can be instructive for a given species, isoform composition or biochemical modification. Here, we describe two methods that visualize microtubule dynamics at high speed and high contrast: (1) Total internal reflection fluorescence microscopy (TIRFM) and (2) label-free interference reflection microscopy (IRM).

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